本文主要研究内容
作者朱俊（2019）在《β-七叶皂苷对结肠癌细胞的抑制作用及分子机制的初探》一文中研究指出：癌症是当今最难攻克的疾病之一。作为恶性肿瘤,结肠癌是一种进展迅速、预后很差的致命性疾病,如何防治结肠癌是医学领域的重要课题之一。中药是中华民族的瑰宝,β-七叶皂苷是从七叶树科天师栗(Aesculus Wilsonii Rebd)或中药娑罗子(Chinese Semen Aesculi)中提取的天然三萜糖苷类化合物。它主要用于脑水肿、抗水肿、增强静脉张力、改善微循环以及慢性静脉功能不全等疾病的治疗,具有血管调节、抑制胃排空、对保护小鼠实验性肝损伤、保护神经和抗肿瘤作用。β-七叶皂苷对常见的大肠癌HT-29细胞的抗癌作用有所报道,但对结肠腺癌Lo Vo细胞的抑制作用及其分子机制,却少见报道。我们利用MTT法、免疫荧光分析、流式细胞术(FCW)、蛋白质印迹法(WB)、激光扫描共聚焦显微镜成像(LSCM)等技术研究β-七叶皂苷抗结肠癌作用及其Wnt/β-catenin信号转导通路的影响。研究β-七叶皂苷对结肠癌细胞的作用,主要从三个方面来阐述:β-七叶皂苷对Lo Vo细胞生物生长抑制作用、β-七叶皂苷对结肠癌Lo Vo细胞的周期和凋亡的影响、核心分子β-catenin的表达水平和细胞内分布。一是β-七叶皂苷对Lo Vo细胞生物生长抑制作用。定量方面,利用MTT法,用0、10μg/ml、30μg/ml、50μg/ml和100μg/ml的β-七叶皂苷处理细胞24h、48h和72h,MTT法检测显示,细胞具有明显的浓度(剂量)和时间相关性,作用浓度越大、作用时间越长,其抑制作用越明显。同时,测得IC50为44.1μg/ml。定性方面,利用间接免疫荧光法,用0、10μg/ml、30μg/ml、50μg/ml和100μg/ml的β-七叶皂苷处理细胞72h,荧光显微镜显示,细胞内的荧光强度随着时间的推移逐渐降低。表明随着药物作用浓度的增高,β-七叶皂苷对Lo Vo细胞生长抑制作用也不断增强,其增殖活性明显被抑制,并且也呈作用-剂量相关性。二是β-七叶皂苷对结肠癌Lo Vo细胞的周期和凋亡的影响。宏观方面,从细胞生物学水平,以一定浓度(50μg/ml)的β-七叶皂苷处理细胞0、24h、48h和72h后,经PI染色于光学显微镜下观察细胞形态学改变,明显逐渐出现核固缩、膜皱缩成泡状、质壁分离等细胞凋亡现象。微观方面,从分子生物学水平,利用DNA含量分析Lo Vo细胞的周期和细胞凋亡,以不同浓度(0、10μg/ml、30μg/ml、50μg/ml和100μg/ml)的β-七叶皂苷处理细胞72h,经PI染色后以流式细胞仪检测细胞DNA含量。结果表明,Lo Vo细胞的周期变短,S期被阻滞,导致进入G2/M期的细胞减少。出现细胞凋亡小峰,细胞凋亡期明显逐渐延长,细胞被β-七叶皂苷诱导凋亡。三是研究β-七叶皂苷对Wnt/β-catenin信号通路核心分子β-catenin表达水平和细胞内分布的影响。蛋白印迹(WB)分析表明药物浓度较低时(10μg/ml和30μg/ml),β-catenin蛋白表达/抑制作用不明显;药物浓度较高时(50μg/ml和100μg/ml),β-catenin蛋白表达/抑制比分别为0.53和0.27,存在显著差异,具有统计学意义(P<0.05)。结合激光扫描共聚焦显微镜(LSCM),可见β-catenin的荧光主要分布在细胞浆(细胞质、细胞器)和细胞膜中,尤其是细胞膜连接部位分布密集,而细胞核中荧光呈多团块状分布,其荧光减弱、变暗,逐渐减少。结果表明,细胞核是影响肿瘤细胞DNA蛋白表达的主要通路。总之,β-七叶皂苷对结肠癌Lo Vo细胞生长具有明显的抑制作用;可以阻滞Lo Vo细胞周期和诱导细胞凋亡;Wnt/β-catenin通路在β-七叶皂苷抗结肠癌机制中发挥了重要作用。
Abstract
ai zheng shi dang jin zui nan gong ke de ji bing zhi yi 。zuo wei e xing zhong liu ,jie chang ai shi yi chong jin zhan xun su 、yu hou hen cha de zhi ming xing ji bing ,ru he fang zhi jie chang ai shi yi xue ling yu de chong yao ke ti zhi yi 。zhong yao shi zhong hua min zu de gui bao ,β-qi xie zao gan shi cong qi xie shu ke tian shi li (Aesculus Wilsonii Rebd)huo zhong yao suo luo zi (Chinese Semen Aesculi)zhong di qu de tian ran san tie tang gan lei hua ge wu 。ta zhu yao yong yu nao shui zhong 、kang shui zhong 、zeng jiang jing mai zhang li 、gai shan wei xun huan yi ji man xing jing mai gong neng bu quan deng ji bing de zhi liao ,ju you xie guan diao jie 、yi zhi wei pai kong 、dui bao hu xiao shu shi yan xing gan sun shang 、bao hu shen jing he kang zhong liu zuo yong 。β-qi xie zao gan dui chang jian de da chang ai HT-29xi bao de kang ai zuo yong you suo bao dao ,dan dui jie chang xian ai Lo Voxi bao de yi zhi zuo yong ji ji fen zi ji zhi ,que shao jian bao dao 。wo men li yong MTTfa 、mian yi ying guang fen xi 、liu shi xi bao shu (FCW)、dan bai zhi yin ji fa (WB)、ji guang sao miao gong ju jiao xian wei jing cheng xiang (LSCM)deng ji shu yan jiu β-qi xie zao gan kang jie chang ai zuo yong ji ji Wnt/β-cateninxin hao zhuai dao tong lu de ying xiang 。yan jiu β-qi xie zao gan dui jie chang ai xi bao de zuo yong ,zhu yao cong san ge fang mian lai chan shu :β-qi xie zao gan dui Lo Voxi bao sheng wu sheng chang yi zhi zuo yong 、β-qi xie zao gan dui jie chang ai Lo Voxi bao de zhou ji he diao wang de ying xiang 、he xin fen zi β-cateninde biao da shui ping he xi bao nei fen bu 。yi shi β-qi xie zao gan dui Lo Voxi bao sheng wu sheng chang yi zhi zuo yong 。ding liang fang mian ,li yong MTTfa ,yong 0、10μg/ml、30μg/ml、50μg/mlhe 100μg/mlde β-qi xie zao gan chu li xi bao 24h、48hhe 72h,MTTfa jian ce xian shi ,xi bao ju you ming xian de nong du (ji liang )he shi jian xiang guan xing ,zuo yong nong du yue da 、zuo yong shi jian yue chang ,ji yi zhi zuo yong yue ming xian 。tong shi ,ce de IC50wei 44.1μg/ml。ding xing fang mian ,li yong jian jie mian yi ying guang fa ,yong 0、10μg/ml、30μg/ml、50μg/mlhe 100μg/mlde β-qi xie zao gan chu li xi bao 72h,ying guang xian wei jing xian shi ,xi bao nei de ying guang jiang du sui zhao shi jian de tui yi zhu jian jiang di 。biao ming sui zhao yao wu zuo yong nong du de zeng gao ,β-qi xie zao gan dui Lo Voxi bao sheng chang yi zhi zuo yong ye bu duan zeng jiang ,ji zeng shi huo xing ming xian bei yi zhi ,bing ju ye cheng zuo yong -ji liang xiang guan xing 。er shi β-qi xie zao gan dui jie chang ai Lo Voxi bao de zhou ji he diao wang de ying xiang 。hong guan fang mian ,cong xi bao sheng wu xue shui ping ,yi yi ding nong du (50μg/ml)de β-qi xie zao gan chu li xi bao 0、24h、48hhe 72hhou ,jing PIran se yu guang xue xian wei jing xia guan cha xi bao xing tai xue gai bian ,ming xian zhu jian chu xian he gu su 、mo zhou su cheng pao zhuang 、zhi bi fen li deng xi bao diao wang xian xiang 。wei guan fang mian ,cong fen zi sheng wu xue shui ping ,li yong DNAhan liang fen xi Lo Voxi bao de zhou ji he xi bao diao wang ,yi bu tong nong du (0、10μg/ml、30μg/ml、50μg/mlhe 100μg/ml)de β-qi xie zao gan chu li xi bao 72h,jing PIran se hou yi liu shi xi bao yi jian ce xi bao DNAhan liang 。jie guo biao ming ,Lo Voxi bao de zhou ji bian duan ,Sji bei zu zhi ,dao zhi jin ru G2/Mji de xi bao jian shao 。chu xian xi bao diao wang xiao feng ,xi bao diao wang ji ming xian zhu jian yan chang ,xi bao bei β-qi xie zao gan you dao diao wang 。san shi yan jiu β-qi xie zao gan dui Wnt/β-cateninxin hao tong lu he xin fen zi β-cateninbiao da shui ping he xi bao nei fen bu de ying xiang 。dan bai yin ji (WB)fen xi biao ming yao wu nong du jiao di shi (10μg/mlhe 30μg/ml),β-catenindan bai biao da /yi zhi zuo yong bu ming xian ;yao wu nong du jiao gao shi (50μg/mlhe 100μg/ml),β-catenindan bai biao da /yi zhi bi fen bie wei 0.53he 0.27,cun zai xian zhe cha yi ,ju you tong ji xue yi yi (P<0.05)。jie ge ji guang sao miao gong ju jiao xian wei jing (LSCM),ke jian β-cateninde ying guang zhu yao fen bu zai xi bao jiang (xi bao zhi 、xi bao qi )he xi bao mo zhong ,you ji shi xi bao mo lian jie bu wei fen bu mi ji ,er xi bao he zhong ying guang cheng duo tuan kuai zhuang fen bu ,ji ying guang jian ruo 、bian an ,zhu jian jian shao 。jie guo biao ming ,xi bao he shi ying xiang zhong liu xi bao DNAdan bai biao da de zhu yao tong lu 。zong zhi ,β-qi xie zao gan dui jie chang ai Lo Voxi bao sheng chang ju you ming xian de yi zhi zuo yong ;ke yi zu zhi Lo Voxi bao zhou ji he you dao xi bao diao wang ;Wnt/β-catenintong lu zai β-qi xie zao gan kang jie chang ai ji zhi zhong fa hui le chong yao zuo yong 。
论文参考文献
论文详细介绍
论文作者分别是来自吉林大学的朱俊，发表于刊物吉林大学2019-06-25论文，是一篇关于七叶皂苷论文,结肠癌论文,细胞论文,抑制作用论文,分子机制论文,吉林大学2019-06-25论文的文章。本文可供学术参考使用，各位学者可以免费参考阅读下载，文章观点不代表本站观点，资料来自吉林大学2019-06-25论文网站，若本站收录的文献无意侵犯了您的著作版权，请联系我们删除。